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TitleStructural basis for the reaction cycle of DASS dicarboxylate transporters.
Journal, issue, pagesElife, Vol. 9, Year 2020
Publish dateSep 1, 2020
AuthorsDavid B Sauer / Noah Trebesch / Jennifer J Marden / Nicolette Cocco / Jinmei Song / Akiko Koide / Shohei Koide / Emad Tajkhorshid / Da-Neng Wang /
PubMed AbstractCitrate, α-ketoglutarate and succinate are TCA cycle intermediates that also play essential roles in metabolic signaling and cellular regulation. These di- and tricarboxylates are imported into the ...Citrate, α-ketoglutarate and succinate are TCA cycle intermediates that also play essential roles in metabolic signaling and cellular regulation. These di- and tricarboxylates are imported into the cell by the divalent anion sodium symporter (DASS) family of plasma membrane transporters, which contains both cotransporters and exchangers. While DASS proteins transport substrates via an elevator mechanism, to date structures are only available for a single DASS cotransporter protein in a substrate-bound, inward-facing state. We report multiple cryo-EM and X-ray structures in four different states, including three hitherto unseen states, along with molecular dynamics simulations, of both a cotransporter and an exchanger. Comparison of these outward- and inward-facing structures reveal how the transport domain translates and rotates within the framework of the scaffold domain through the transport cycle. Additionally, we propose that DASS transporters ensure substrate coupling by a charge-compensation mechanism, and by structural changes upon substrate release.
External linksElife / PubMed:32869741 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2.86 - 3.92 Å
Structure data

EMDB-21902, PDB-6wu1:
Structure of apo LaINDY
Method: EM (single particle) / Resolution: 3.09 Å

EMDB-21903, PDB-6wu2:
Structure of the LaINDY-malate complex
Method: EM (single particle) / Resolution: 3.36 Å

EMDB-21904, PDB-6wu3:
Structure of VcINDY-Na+ in amphipol
Method: EM (single particle) / Resolution: 3.16 Å

EMDB-21905, PDB-6wu4:
Structure of the LaINDY-alpha-ketoglutarate complex
Method: EM (single particle) / Resolution: 3.71 Å

EMDB-21928, PDB-6ww5:
Structure of VcINDY-Na-Fab84 in nanodisc
Method: EM (single particle) / Resolution: 3.15 Å

PDB-6wtw:
Structure of LaINDY crystallized in the presence of alpha-ketoglutarate and malate
Method: X-RAY DIFFRACTION / Resolution: 2.86 Å

PDB-6wtx:
Structure of VcINDY in complex with terephthalate
Method: X-RAY DIFFRACTION / Resolution: 3.92 Å

Chemicals

ChemComp-UB7:
terephthalic acid / Terephthalic acid

ChemComp-NA:
SODIUM ION / Sodium

ChemComp-HEX:
HEXANE / Hexane

ChemComp-D10:
DECANE / Decane

ChemComp-OCT:
N-OCTANE / Octane

ChemComp-6PE:
1,2-DIHEXANOYL-SN-GLYCERO-3-PHOSPHOETHANOLAMINE

Source
  • lactobacillus acidophilus (bacteria)
  • vibrio cholerae (bacteria)
  • homo sapiens (human)
KeywordsTRANSPORT PROTEIN / Transporter / Structural Genomics / PSI-Biology / New York Consortium on Membrane Protein Structure / NYCOMPS / MEMBRANE PROTEIN

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